小鼠PD-L1真核表达载体的构建及其在CHO细胞中的稳定表达  

Construction of eukaryotic expressing vector of mouse PD-L1 and establishment of stable transfectant CHO cell line

作　　者：付文哲[1] 陈永井[1] 王勤[1] 张雪琨[1] 马钰[1] 张学光[1] 

机构地区：[1]苏州大学生物技术研究所,江苏苏州215007

出　　处：《苏州大学学报（医学版）》2012年第2期224-227,共4页Suzhou University Journal of Medical Science

摘　　要：目的构建含有小鼠PD-L1基因的真核表达载体,并通过转染获得稳定表达小鼠PD-L1分子的CHO细胞系。方法从小鼠脾细胞总RNA逆转录的cDNA中扩增出PD-L1基因,通过双酶切(Xho I和EcoR I)装入真核表达载体pIRES2-EGFP中,脂质体法转染CHO细胞,经G418筛选后,建立稳定高表达PD-L1分子的CHO细胞系。结果构建了真核表达载体pIRES2-EGFP/PD-L1,建立了稳定表达PD-L1目的基因的CHO细胞系。结论成功构建了PD-L1真核表达载体并获得了稳定表达该分子的CHO细胞系为后续研究奠定了物质基础。Objective To construct eukaryotic expressing vector of mouse PD-L1 gene and transfect CHO cells so as to establish stable CHO cell line expressing PD-L1. Methods Mouse PD-L1 gene frag- ment was obtained by RT-PCR, recombinant vector was constructed by linking the PCR products and pIRES2-EGFP which was digested by Xho I and EcoR I. Then pIRES2-EGFP/PD-LI was transfected into CHO cells by Lipofectamine 2 000. Results The eukaryotic expressing vector pIRES2-EGFP/PD-L1 was constructed. Stable transfected CHO cell line expressing pD-L1 gene was established. Conclusion The construction of eukaryotic expressing vector pIRES2-EGFP/PD-L1 and the establishment of stable trans- fected CHO cell line have provided solid experimental foundation for further studies.

关 键 词：PD-L1 真核表达载体 转染 CHO细胞 

分 类 号：R394.3[医药卫生—医学遗传学] Q75[医药卫生—基础医学]